Abstract: The partition of human antibodies in aqueous two-phase systems (ATPSs) of polyethylene glycol (PEG) and phosphate was systematically studied using first pure proteins systems and then an artificial mixture of proteins containing 1mg/ml human immunoglobulin G (IgG), 10mg/ml serum albumin and 2mg/ml myoglobin. Preliminary results obtained using pure proteins systems indicated that the PEG molecular weight and concentration, the pH value and the salts concentration had a pronounced effect on the partitioning behaviour of all proteins. For high ionic strengths and pH values higher than the isoelectric point (pI) of the contaminant proteins, IgG could be selectively recovered on the top phase. According to these results, a face centred composite design was performed in order to optimise the purification of IgG from the mixture of proteins. The optimal conditions for the isolation of IgG were observed for high concentrations of NaCl and low concentrations of both phase forming components. The best purification was achieved using an ATPS containing 8% (w/w) PEG 3350, 10% (w/w) phosphate pH 6 and 15% (w/w) NaCl. A recovery yield of 101+/-7%, a purity of 99+/-0% and a yield of native IgG of 97+/-4% were obtained. Back extraction studies of IgG to a new phosphate phase were performed and higher yields were obtained using 10% phosphate buffer at pH 6. The total extraction yield was 76% and the purity 100%.

Abstract: OBJECTIVE: To assess the effects of a single intravenous dose of butorphanol (0.1 mg kg(-1)) on the nociceptive withdrawal reflex (NWR) using threshold, suprathreshold and repeated subthreshold electrical stimuli in conscious horses. STUDY DESIGN: 'Unblinded', prospective experimental study. ANIMALS: Ten adult horses, five geldings and five mares, mean body mass 517 kg (range 487-569 kg). METHODS: The NWR was elicited using single transcutaneous electrical stimulation of the palmar digital nerve. Repeated stimulations were applied to evoke temporal summation. Surface electromyography was performed to record and quantify the responses of the common digital extensor muscle to stimulation and behavioural reactions were scored. Before butorphanol administration and at fixed time points up to 2 hours after injection, baseline threshold intensities for NWR and temporal summation were defined and single suprathreshold stimulations applied. Friedman repeated-measures analysis of variance on ranks and Wilcoxon signed-rank test were used with the Student-Newman-Keul's method applied post-hoc. The level of significance (alpha) was set at 0.05. RESULTS: Butorphanol did not modify either the thresholds for NWR and temporal summation or the reaction scores, but the difference between suprathreshold and threshold reflex amplitudes was reduced when single stimulation was applied. Upon repeated stimulation after butorphanol administration, a significant decrease in the relative amplitude was calculated for both the 30-80 and the 80-200 millisecond intervals after each stimulus, and for the whole post-stimulation interval in the right thoracic limb. In the left thoracic limb a decrease in the relative amplitude was found only in the 30-80 millisecond epoch. CONCLUSION: Butorphanol at 0.1 mg kg(-1) has no direct action on spinal Adelta nociceptive activity but may have some supraspinal effects that reduce the gain of the nociceptive system. CLINICAL RELEVANCE: Butorphanol has minimal effect on sharp immediate Adelta-mediated pain but may alter spinal processing and decrease the delayed sensations of pain.

Abstract: The ovine arena test in conjunction with multivariate statistical analysis (factor analysis) may be a means of measuring ovine temperament for practical purposes. Stability of factor construction over time and between groups of animals is considered to demonstrate trait consistency and is, therefore, one of the first steps in validating a temperament/personality test from this perspective. The aim of this study, therefore, was to assess the stability of factor construction, as a measure of trait consistency, using arena test data from three groups of animals with one group (Group 1) tested repeatedly over three rounds (twice at 8 months and once at 22 months of age). Group 1 consisted of 193 mule (Bluefaced Leicester Sire x Scottish Blackface/Welsh Speckled Face dam), ewe lambs (8 months old). Groups 2 and 3 consisted of 189 and 185 mules, respectively (14 months old). All animals were tested for 6 min in a 13 m x 3 m arena. Factor analysis (varimax rotation) was performed twice on the behavioural data (latency to bleat, total number of vocalisations, distance travelled, time spent in different areas of the arena and number of times crossing in and out of pertinent areas), initially using all data recorded on a per minute basis (`Per Minute') for all 6 min of the test (10 factors extracted) and then using total values (`Total'), the summation of the 6 min for each behaviour measured (4 factors extracted). Stability of factor loadings between rounds and between groups was tested using Kendall's coefficient of concordance. For the `Per Minute' data, 5 out of the 10 factors showed significant (p < 0.05) concordance between rounds whilst 9 out of 10 factors showed significant (p < 0.05) concordance between groups. All four factors generated from the `Total' data demonstrated significant (p < 0.05) concordance between rounds and between groups. The four factors generated from the `Total' data were considered to be of potential merit for future studies. These factors were named--`conspecific motivation-fear', `conspecific motivation-distress', `activity' and `low conspecific motivation'.